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Related Experiment Video

Updated: May 28, 2026

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

Kinase inhibitor selectivity profiling using differential scanning fluorimetry.

Oleg Fedorov1, Frank H Niesen, Stefan Knapp

  • 1Structural Genomics Consortium, Oxford University, Oxford, UK. oleg.fedorov@sgc.ox.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|October 1, 2011
PubMed
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Differential scanning fluorimetry (DSF) offers a fast, robust method for screening protein kinase inhibitors. This technique monitors thermal stabilization upon ligand binding, enabling selectivity profiling without needing enzyme activity data.

Area of Science:

  • Biochemistry
  • Drug Discovery
  • Biophysics

Background:

  • Enzymatic assays are standard for screening protein kinase inhibitors but require active enzymes, substrates, and optimization.
  • Alternative ligand-binding assays are emerging to overcome limitations of traditional enzymatic methods.

Purpose of the Study:

  • To present differential scanning fluorimetry (DSF) as a fast, robust, and inexpensive method for screening inhibitor libraries against protein kinases.
  • To provide an experimental protocol for applying DSF in drug discovery.

Main Methods:

  • Differential scanning fluorimetry (DSF) was employed to monitor the thermal stabilization of proteins upon ligand binding.
  • The method was applied to screen medium-sized inhibitor libraries (<10,000 compounds) against protein kinases.

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Last Updated: May 28, 2026

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery
06:26

Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery

Published on: May 16, 2021

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

  • Selectivity profiling was performed without prior knowledge of enzyme activity or substrate.

    • Main Results:

    • DSF provides a rapid and reproducible approach for inhibitor screening.
    • The method allows for selectivity profiling of protein kinases.
    • DSF data showed high reproducibility and good correlation with other ligand-binding assays and enzymatic assays.

    Conclusions:

    • DSF is a valuable tool for fast and robust screening of protein kinase inhibitors.
    • This method offers an alternative to traditional enzymatic assays, simplifying the screening process.
    • DSF enables efficient selectivity profiling, aiding in drug discovery efforts.